Noxic effects of polystyrene microparticles on murine macrophages and epithelial cells

Abstract Microplastic (MP) contamination has been identified as an ecological problem with an increasing impact on everyday life. Yet, possible effects of MP at the cellular level are still poorly understood. Here, the interaction of murine macrophages (J774A.1, ImKC) and epithelial cells (STC-1, BNL CL.2) with well-characterized poly(styrene) MP particles (MPP) of varying sizes (0.2–6.0 µm) was studied. Macrophages are expected to actively engulf particles which could be confirmed in this study, while epithelial cells are found in tissues with direct contact with ingested or inhaled MPP. Here, the epithelial cells from both investigated cell lines did not ingest MPP in significant numbers. Concomitantly, no cytotoxic effects nor any influence on cellular proliferation were observed. Cells from the two macrophage cell lines showed high ingestion of MPP of all sizes, but cytotoxic effects were observed only for one of them (ImKC) and only at MPP concentrations above 250 µg/mL. Indications of cellular stress as well as effects on cell proliferation were observed for cell populations with high particle cell interactions

Generation of Recombinant Primary Human B Lymphocytes Using Non-Viral Vectors

Although the development of gene delivery systems based on non-viral vectors is advancing, it remains a challenge to deliver plasmid DNA into human blood cells. The current “gold standard”, namely linear polyethyleneimine (l-PEI 25 kDa), in particular, is unable to produce transgene expression levels >5% in primary human B lymphocytes. Here, it is demonstrated that a well-defined 24-armed poly(2-dimethylamino) ethyl methacrylate (PDMAEMA, 755 kDa) nano-star is able to reproducibly elicit high transgene expression (40%) at sufficient residual viability (69%) in primary human B cells derived from tonsillar tissue. Moreover, our results indicate that the length of the mitogenic stimulation prior to transfection is an important parameter that must be established during the development of the transfection protocol. In our hands, four days of stimulation with rhCD40L post-thawing led to the best transfection results in terms of TE and cell survival. Most importantly, our data argue for an impact of the B cell subsets on the transfection outcomes, underlining that the complexity and heterogeneity of a given B cell population pre- and post-transfection is a critical parameter to consider in the multiparametric approach required for the implementation of the transfection protocol

The Kolmogorov-Smirnov test for the CMB

We investigate the statistics of the cosmic microwave background using the Kolmogorov-Smirnov test. We show that, when we correctly de-correlate the data, the partition function of the Kolmogorov stochasticity parameter is compatible with the Kolmogorov distribution and, contrary to previous claims, the CMB data are compatible with Gaussian fluctuations with the correlation function given by standard Lambda-CDM. We then use the Kolmogorov-Smirnov test to derive upper bounds on residual point source power in the CMB, and indicate the promise of this statistics for further datasets, especially Planck, to search for deviations from Gaussianity and for detecting point sources and Galactic foregrounds.Comment: Improved significance of the results (which remain unchanged) by using patches instead of ring segments in the analysis. Added sky maps of the Kolmogorov-parameter for original and de-correlated CMB ma

<i>Malmgrenia louiseae</i> sp. nov., a new scale worm species (Polychaeta: Polynoidae) from southern Europe with a key to European <i>Malmgrenia</i> species

International audienceMalmgrenia louiseae sp. nov. is described from both the Western Mediterranean in the Gulf of Lions, and the Northeast Atlantic from off Portugal and the Bay of Biscay. The species was found in muddy sediments in shallow water and is possibly associated with echiurids or synaptid holothurians. Malmgrenia louiseae sp. nov. can be clearly distinguished from all other known Malmgrenia species by the presence of an infra-acicular process in addition to the supra-acicular process on the acicular lobe of the neuropodia, the lack of microtubercules on the elytra, two kinds of notochaetae (stout with blunt tip and slender with fine pointed tip), and exclusively unidentate neurochaetae. An identification key to the Northeast Atlantic and Mediterranean Malmgrenia species is provided

In vitro cultivation of primary intestinal cells from Eisenia fetida as basis for ecotoxicological studies

The earthworm Eisenia fetida is a commonly used model organism for unspecific soil feeders in ecotoxicological studies. Its intestinal cells are the first to encounter possible pollutants co-ingested by the earthworm, which makes them prime candidates for studies of toxic effects of environmental pollutants on the cellular as compared to the organismic level. In this context, the aim of this study was to demonstrate the suitability of preparations of primary intestinal E. fetida cells for in vitro ecotoxicological studies. For this purpose, a suitable isolation and cultivation protocol was established. Cells were isolated directly from the intestine, maintaining >85% viability during subsequent cultivations (up to 144 h). Exposure to established pollutants and soil elutriates comprising silver nanoparticles and metal ions (Cu(2+), Cd(2+)) induced a significant decrease in the metabolic activity of the cells. In case of microplastic particles (MP particles), namely 0.2, 0.5, 2.0, and 3.0 µm diameter polystyrene (PS) beads as well as 0.5 and 2.0 µm diameter polylactic acid (PLA) beads, no active uptake was observed. Slight positive as well as negative dose and size dependent effects on the metabolism were seen, which to some extent might correlate with effects on the organismic level